Environ Monit Assess (2010) 161:473–483
DOI 10.1007/s10661-009-0761-8
Assessment of microbiological indoor air quality
in an Italian office building equipped
with an HVAC system
Sa. Bonetta · Si. Bonetta · S. Mosso ·
S. Sampò · E. Carraro
Received: 25 July 2008 / Accepted: 27 January 2009 / Published online: 18 February 2009
© Springer Science + Business Media B.V. 2009
Abstract The purpose of this study was to eval-
uate the level and composition of bacteria and
fungi in the indoor air of an Italian office building
equipped with a heating, ventilation and air con-
ditioning (HVAC) system. Airborne bacteria and
fungi were collected in three open-space offices
during different seasons. The microbial levels in
the outdoor air, supply air diffusers, fan coil air
flow and air treatment unit humidification water
tank were used to evaluate the influence of the
HVAC system on indoor air quality (IAQ). A
medium–low level of bacterial contamination (50–
500 CFU/m3 ) was found in indoor air. Staphylo-
coccus and Micrococcus were the most commonly
found genera, probably due to human presence.
Electronic supplementary material The online version
of this article (doi:10.1007/s10661-009-0761-8) contains
supplementary material, which is available to
authorized users.
Sa. Bonetta · Si. Bonetta · S. Sampò · E. Carraro (B)
Dipartimento di Scienze dell’Ambiente e della Vita,
University of Piemonte Orientale “Amedeo
Avogadro”, Via Bellini 25/G,
15100 Alessandria, Italy
e-mail: elisabetta.carraro@mfn.unipmn.it
S. Mosso
Telecom Italia Laboratories,
Via G. Reiss Romoli 274, 10148 Turin, Italy
A high fungal concentration was measured due
to a flood that occurred during the winter. The
indoor seasonal distribution of fungal genera was
related to the fungal outdoor distribution. Sig-
nificant seasonal and daily variation in airborne
microorganisms was found, underlining a relation-
ship with the frequency of HVAC system switch-
ing on/off. The results of this monitoring highlight
the role of the HVAC system on IAQ and could
be useful to better characterise bacterial and fun-
gal population in the indoor air of office buildings.
Keywords Bacteria · Fungi · Indoor air ·
HVAC system · Office building
Introduction
Indoor air quality (IAQ) is an increasingly im-
portant issue for occupational and public health
(Clarke and Nikkel 1995; Reynolds et al. 2001).
Health effects related to IAQ have increased,
perhaps due to some energy-saving measures pro-
vided for buildings such as tight sealing (Jones
1999), air recirculation and the introduction of
heating ventilation air conditioning (HVAC) sys-
tems (Nathanson 1995). In addition, an ageing
population, an increasing number of sensitive in-
dividuals and a tendency to spend more time
indoors (90% of life time) further worsen this
problem (Tringe et al. 2008). Although discussions
474
about indoor air contamination frequently con-
centrate on chemical pollutants, the health effects
of inhaled biological particles should not be over-
looked, as a large variety of biological material is
present in indoor environment (Montanaro 1997).
Bioaerosols are airborne particles that are ei-
ther living (e.g. bacteria, fungi) or originate from
living organisms that are ubiquitous, highly vari-
able and complex and are natural or man-made
in origin. The sampling and analysis of airborne
microorganisms in indoor air has received atten-
tion in recent years (Kim and Kim 2007; Huttunen
et al. 2008; Stanley et al. 2008). Bioaerosols con-
tribute to about 5–34% of indoor air pollution
(Srikanth et al. 2008). The source of bioaerosols
in indoor air include furnishing and building ma-
terials, microbiological contamination within the
walls, and ceiling and floor cavities. Another sig-
nificant source of airborne bacteria in the indoor
environment is the occupants (Loftness et al. 2007;
Stanley et al. 2008). Many indoor bioaerosols orig-
inate outdoors through the opening of doors and
windows or through cracks in the building enve-
lope. However, one of the most important factors
affecting IAQ is how the building is heated, ven-
tilated and air conditioned (Seppänen and Fisk
2002), considering that in many cases, particularly
in office buildings, these functions are integrated
into an HVAC system.
Although HVAC systems can help remove and/
or dilute more than 80% of aerosols from the out-
doors, they can also provide favourable breeding
grounds for bioaerosols to colonise (Law et al.
2001). A microbiological growth may occur in
an HVAC system equipped with low-efficiency
filters, humidifiers that use water recycling or in
areas in which water condensation remains stag-
nant and large recirculation of the air is present
(Wu et al. 2005; Huang et al. 2008). Microor-
ganisms can thus spread in the indoor air by
the HVAC system and be inhaled by the people
working or living in buildings (Parat et al. 1997;
Mendell et al. 2008).
Numerous studies published during the past
10–15 years have produced rather solid scien-
tific evidence that indoor aerosol particles, es-
pecially in the respirable fraction, are associated
with health effects. Viable bioaerosol particles,
including viruses, bacteria and fungi, have been
Environ Monit Assess (2010) 161:473–483
associated with respiratory allergies and asthma
and have been linked to the airborne transmission
of various infections known as building-related
illnesses (e.g. Legionnaires’ disease, aspergillosis;
Stetzenbach 1997; Huang et al. 2008). Airborne
bacteria and fungi can also be the cause of non-
specific diseases such as sick building syndrome
(SBS). Numerous scientific studies have docu-
mented that SBS is surprisingly common even in
buildings without widespread health complaints
(Mendell et al. 2008). Moreover, epidemiological
investigations showed that SBS and hypersensitiv-
ity diseases (humidifier fever or asthma) are often
associated with high airborne microbial concen-
tration exposure (ACGIH 1999; Shoemaker and
House 2006).
To contribute to the knowledge on IAQ, this
study evaluates the level and composition of bac-
terial and fungal contamination of indoor air in
an Italian office building equipped with an HVAC
system during different seasons. The influence of
the HVAC characteristics and human activities on
IAQ was also investigated.
Materials and methods
Description of the site
The study was carried out in an office building
(35 years old) with a central HVAC system and
open-space offices located in a busy and subur-
ban zone of Turin (Piedmont, Italy). There were
two or three occupants per office, computers and
printers. The HVAC system was composed of the
following: an intake for outdoor air located on the
roof of the building, vent ducts at the ceiling for
air removal, and a duct bringing outdoor air to
the air treatment unit (ATU, 4 years old). In the
ATU, fresh (50% minimum) and recycled air were
mixed and filtered (85% removal for ≥3 μm parti-
cles prefilter according to ASHRAE 52–76 with a
bag-type filter), heated or cooled, dehumidified or
humidified (air humidification tank) and distrib-
uted through a duct network to vent ducts placed
in the false ceiling. Heating and cooling devices
(fan coil units) were distributed in open spaces
to heat or cool indoor recycled air. The windows
were locked, and building occupants could only
Environ Monit Assess (2010) 161:473–483
regulate the fan coil units by switching them on
or off.
Sampling
A diagram of the sampling points is shown in
Fig. 1. Three open-space offices (A, B and C) were
selected to follow the air duct distribution from
the fresh air intake to the end. Air was sampled
for microbiological analysis in the centre of each
open-space office, at 1.5 m from the floor (at the
workstation level) and was collected twice a day
(at 9.00 a.m. and 4.30 p.m.), three times a week
(Monday, Wednesday and Friday) and 1 week
during different seasons (winter, spring, summer).
Since an unusual flood occurred during the winter,
another sampling was repeated during the follow-
ing winter (winter2).
To evaluate the HVAC influence on in-
door microbiological contamination, additional
evaluations were performed once a week (on
Wednesday) for each season. In particular, the
bacterial and fungal counts at the intake point
of the ATU system (Fig. 1) and at the level of
the two ventilation shafts in the indoor air (the
first and the last ventilation shafts after the ATU
system) were analysed. Microbiological analyses
were also performed on the ATU humidification
water tank. The air flow of the fan coil units was
also sampled in the same offices in which IAQ
was monitored to evaluate its influence on air
contamination. A total of 240 air samples were
collected for bacterial and fungal measurements:
Fig. 1 Diagram of the INTAKE POINT o
sampling points in
the office building
FILTER
w UTA
WATER TANK
S1
FIRST SHAFT
O: outdoor air;
S1: first ventilation shaft;
S2: last ventilation shaft;
W: humidification water tank;
A,B,C: offices;
a,b,c: fan coil units.
475
(three parameters: bacteria at 22◦ C, bacteria at
37◦ C and fungi) × (three offices: A, B and C) ×
(two daily samplings: a.m./p.m.) × (three times a
week: Monday, Wednesday and Friday) × (four
seasons: winter, spring, summer, winter2) + [(one
outdoor sample + two ventilation shafts + three
fan coil units) × (four seasons: winter, spring,
summer, winter2)].
Airborne bacteria and fungi were collected
using a calibrated impactor sampler (SAS Super
180™, International PBI, Milan) at an air flow rate
of 180 L/min (sampling time 1.10 and 1.20 min
for fungal and bacterial samplings, respectively).
Three plates per each sampling point were taken
to evaluate the different microbiological parame-
ters and quality assurance/quality control aspect.
Field blank plates for quality control (a mini-
mum of one for each medium) accompanied the
sampler during the transport and air monitoring.
These field blanks were processed and analysed
exactly as the other plates.
Microbiological analyses
Total bacterial counts were evaluated on trypti-
case soy agar supplemented with cycloheximide
(100 μg/L), and fungal counts were performed
on Rose-Bengal Agar with chloramphenicol
(100 μg/L). Petri dishes were incubated for 24 h
at 37◦ C and 48h at 22◦ C for bacterial counts and
4–8 days at 24◦ C for fungal counts. The mean
value of the triplicate samples was calculated,
and results were expressed in colony forming
OPEN SPACE OFFICES
a b c
A B C
S2
LAST SHAFT
476 Environ Monit Assess (2010) 161:473–483

units per cubic meter (CFU/m3 ), corrected based
on the sampler’s manufacturer instructions. The
consistency of triplicate counts was evaluated us-
ing a χ 2 index to confirm the suitability of the
mean value of the three measurements. The most
widespread bacterial and fungal colonies for each
sampling were selected and isolated from indoor
and outdoor air samples on the basis of their
morphology.
Water samples collected in the air humidifica-
tion tank of the ATU were analysed for Legionella
spp. (ISO 1998) and total bacterial counts (22◦ C
and 37◦ C counts; EN ISO 1999).
Microbial identification
their micro- and macro-morphological character-
istics using standard taxonomic keys (Von Arx
1981).
Statistical analyses
SYSTAT software for Windows, version 8.0, was
used for statistical analysis. Analysis of variance
(ANOVA) was performed on bacteria (22◦ C and
37◦ C counts) and fungi indoor concentrations us-
ing season, day of the working week, and morning
or afternoon as main parameters. Pearson’s corre-
lation coefficient was used to study the relation-
ship among the microbial parameters analysed.

Bacterial identifications by metabolic fingerprint- Results

ing analysis were performed using the Biolog
system (Biolog Inc., Hayward, CA, USA), orig-
inally created for soil and water microorganisms
identification (Avidano et al. 2005). This method
tests the ability of a microorganism to utilise or ox-
idise different carbon sources. Tetrazolium violet
is used as a redox dye. Briefly, bacteria were
grown on Biolog Universal Growth agar at 37◦ C
for 24 h or at 22◦ C for 48 h. Colonies were sus-
pended in a 0.4% saline solution and the inoculum
density adjusted to the specified turbidity range.
The bacterial suspensions were inoculated in a
Biolog GP Microplate and were incubated at 37◦ C
or 22◦ C then manually read after 24 h. The pattern
of purple wells was analysed by Biolog Microlog
2 software.
Fungal genera were identified by colony char-
acteristics and microscopic examination based on
Microbiological indoor air quality
Table 1 summarises the values of bacterial and
fungal indoor contamination during seasonal sam-
plings. For each season, bacterial and fungal
counts detected in the three offices (A, B and C)
on Monday, Wednesday and Friday were pooled
and reported as a weekly mean with minimum and
maximum values. The counts observed were com-
pared with categories of contamination indicated
in the guidelines of the Commission of European
Communities (CEC 1993, Table 2).
The first winter sampling revealed unusually
high indoor fungal concentrations. During this
sampling, high fungal contamination was found
in all offices and a value >2,000 CFU/m3 was
observed in office C (Table 1). This finding is

Table 1 Indoor air bacterial (22◦ C and 37◦ C) and fungal concentrations (CFU/m3 ) during seasonal samplings
Parameter (CFU/m3 ) Winter (n = 18)a Spring (n = 18)a Summer (n = 18)a Winter2 (n = 18)a
Meanb Min Max Meanb Min Max Meanb Min Max Meanb Min Max
Bacteria 37◦ C m 102 32 248 97 36 256 198 120 368 253 116 496
a 76 44 128 65 32 96 136 68 324 158 84 268
Bacteria 22◦ C m 72 32 200 120 56 320 176 112 240 271 128 500
a 28 4 56 129 40 308 134 52 368 160 72 304
Fungi 24◦ C m 514 50 1,528 321 55 465 44 25 90 49 10 100
a 872 30 2,315 43 25 100 44 10 75 39 10 70
m morning, a afternoon, Min minimum value, Max maximum value
a Number of samples analysed for each parameter
b Weekly mean of microbiological counts recovered in the three offices (A, B, and C) on Monday, Wednesday, and Friday
Environ Monit Assess (2010) 161:473–483
Table 2 Categories of CFU/m3 (mixed population of bac-
teria and of fungi) for non-industrial indoor environments
(CEC 1993)
Category Bacteria Fungi
Very low <50 <25
Low <100 <100
Intermediate <500 <500
High <2,000 <2,000
Very high >2,000 >2,000
These categories are based on the range of values obtained
in indoor environments and not on a health risk evaluation
probably related to a flood that occurred in
Turin near the investigated building during sam-
pling. Massive mould growth was also reported
in other studies on flood-damaged homes (Riggs
et al. 2008; Rao et al. 2007). For this reason,
another sampling was repeated the following win-
ter (winter2).
In the other seasonal samplings (spring, sum-
mer and winter2), the fungal concentration de-
creased, and values lower than 500 CFU/m3
(medium–low contamination levels in spring) and
100 CFU/m3 (very low–low contamination levels
in summer and winter2) were observed. The low
contamination reported in winter2 confirms that
the high fungal levels encountered in the first
winter sampling do not represent typical winter
contamination and that they had been influenced
by the flood (Riggs et al. 2008; Rao et al. 2007).
Bacterial concentrations (37◦ C) ranged be-
tween 32 and 496 CFU/m3 (mean 136 CFU/m3 ),
with values lower than 500 CFU/m3 (medium–low
contamination levels). A similar trend was ob-
served for bacteria at 22◦ C, showing a mean con-
centration of 136 CFU/m3 (range 4–500 CFU/m3 ).
The ANOVA test performed on total raw data
(excluding the first winter sampling) revealed sig-
nificant differences in bacterial (22◦ C and 37◦ C)
Table 3 The analysis of variance on bacterial and fungal counts with respect to season, sampling point (office), day of work
week, and sampling hour
Bacteria 22◦ C
Season p < 0.001 (Sp > Su > W2)
Office n.s.
Days of working week p < 0.05 (F > W > M)
Hour p < 0.05 (m > a)
Values obtained excluded the anomalous winter sample
n.s. not significant, Sp spring, Su summer, W2 winter2, F Friday, W Wednesday, M Monday, m morning, a afternoon
477
and fungal indoor contamination related to the
season (Table 3). The bacterial counts (22◦ C and
37◦ C) increased from spring to summer and win-
ter2, while the fungal concentration increased
in the spring. Similar trends were also observed
in other studies (Sen and Asan 2009). Further-
more, a significant difference in daily bacterial
(22◦ C and 37◦ C) and fungal indoor contamina-
tion was detected, showing a trend with higher
contamination levels in the morning than in the
afternoon (Table 3). A significant work week
trend of bacterial indoor contamination (22◦ C) of
Friday > Wednesday > Monday was revealed by
the ANOVA.
No significant relationship among bacterial
concentrations at 22◦ C and 37◦ C and fungal
counts was revealed by the Pearson’s correlation
coefficient.
HVAC system
Air treatment unit
To study the influence of the ATU system on
IAQ, microbial contamination of outdoor and in-
door air sampled near supply air diffusers was
evaluated (Table 4). Fungal and bacterial (22◦ C)
levels from outdoor air increased from winter to
summer, except for the fungal air concentration
measured during the flood, which was unusually
high. As reported by other authors (Pastuszka
et al. 2000; Shelton et al. 2002; Lee et al. 2006),
seasonal high temperatures and humidity can in-
crease microbiological activity, leading to an in-
crease of bacterial and fungal counts.
In the first winter sampling, similar fungal
counts in outdoor and indoor air diffusers (>3,000
CFU/m3 , SAS upper detection limit) were ob-
served, so it is possible that the ATU could not
Bacteria 37◦ C Fungi 24◦ C
p < 0.001 (Sp > Su > W2) p < 0.001 (W2 ≈ Su > Sp)
n.s. n.s.
n.s. n.s.
p < 0.05 (m > a) p < 0.05 (m > a)
478 Environ Monit Assess (2010) 161:473–483

Table 4 Bacterial (22◦ C and 37◦ C) and fungal concentration (CFU/m3 ) in outdoor air and at the ventilation shafts during
seasonal samplings
CFU/m3 Winter Spring Summer Winter2
Meana ± SD Meana ± SD Meana ± SD Meana ± SD
Bacteria 37◦ C O 114 ± 62 96 ± 17 284 ± 86 248 ± 53
Bacteria 22◦ C O 189 ± 17 352 ± 57 600 ± 127 308 ± 87
Fungi 24◦ C O >3,268b 145 ± 15 695 ± 145 115 ± 8
Bacteria 37◦ C S1 20 ± 4 44 ± 5 84 ± 11 132 ± 16
Bacteria 22◦ C S1 4±2 144 ± 39 120 ± 30 228 ± 70
Fungi 24◦ C S1 >3,268b 60 ± 10 70 ± 4 20 ± 8
Bacteria 37◦ C S2 48 ± 5 48 ± 20 76 ± 38 76 ± 40
Bacteria 22◦ C S2 16 ± 1 104 ± 2 60 ± 28 48 ± 11
Fungi 24◦ C S2 >3,268b 30 ± 3 50 ± 18 40 ± 23
SD standard deviation, O outdoor air collected at the intake point of the ATU system, S1 indoor air at the level of the first
ventilation shaft after the ATU system, S2 indoor air at the level of the last ventilation shaft after the ATU system
a Mean value of the three measurements
b SAS upper detection limit calculated, according to the manufacturer, considering the maximum number of colonies in the

Petri dishes counted with the SAS Super 180

remove the high fungal contamination in the out-
door air. Otherwise, in this sampling, outdoor
bacterial values (22◦ C and 37◦ C) were lower than
fungal counts, and the ATU was able to remove
most of it.
During all the other samplings, the microbio-
logical contamination of the air measured at the
ATU air diffusers revealed a lower bacterial and
fungal concentration than outdoors, with values
lower than 500 or 100 CFU/m3 .
Fan coil air
Table 5 summarises bacterial and fungal concen-
trations in the fan coil unit air flow during seasonal
samplings. High fungal contamination (range 500–
2,000 CFU/m3 ) of the fan coil unit air flow was
observed in the first winter sampling when high
fungal counts were found in indoor and outdoor
air. During other samplings, both bacterial and
fungal concentrations were medium–low or some-
times very low (<500 or <50 CFU/m3 ), similar
levels to that observed in the indoor air.
Indoor–outdoor ratio
The indoor/outdoor ratio (I/O; Table 6) was cal-
culated using the indoor (mean values obtained in
the open-space offices on Wednesday) and out-
door microbial counts obtained during the four
samplings. I/O ratios showed that fungal counts
and 22◦ C bacterial concentrations were always
higher outdoors than indoors, in agreement with
other studies performed in office buildings (Burge
et al. 2000). On the other hand, I/O values close
to 1 for bacterial counts at 37◦ C indicated higher

Table 5 Bacterial (22◦ C and 37◦ C) and fungal concentration (CFU/m3 ) in the fan coil unit’s air flow during seasonal
samplings
Parameter (CFU/m3 ) Winter (n = 3)a Spring (n = 3)a Summer (n = 3)a Winter2 (n = 3)a
Meanb Min Max Meanb Min Max Meanb Min Max Meanb Min Max
Bacteria 37◦ C 82 60 113 52 36 80 128 32 200 96 80 112
Bacteria 22◦ C 36 20 47 71 44 116 95 32 128 180 160 200
Fungi 24◦ C 935 560 1,465 15 10 20 50 30 65 60 20 100
Min minimum value, Max maximum value
a Number of samples analysed for each parameter
b Mean of microbiological counts recovered at the three fan coils air flow (office A, B, and C) on Wednesday
Environ Monit Assess (2010) 161:473–483 479

Table 6 Indoor/outdoor ratios calculated for bacterial and

fungal loads in seasonal samplings cus), in the external ear auditory meatus (Staphy-
lococcus auricularis) or on the human scalp
Season Ia /Ob
(Staphylococcus capitis) were also found. Some
Bacteria 22◦ C Bacteria 37◦ C Fungi 24◦ C
of the identified bacteria (Staphylococcus epider-
Winter 0.63 0.82 0.34
Spring 0.39 1.08 0.80
midis, S. haemolyticus, Staphylococcus warneri
Summer 0.80 1.06 0.38 and Staphylococcus hominis) can be considered to
Winter2 0.70 1.13 0.28 be opportunistic pathogens, while others (Kurtia
a Means of microbiological counts obtained at the level of
gibsonii, Kocuria rosea erythromixa and Tsuka-
open-spaces on Wednesday murella inchonensis) are typically found in envi-
b Microbiological counts obtained at the intake point of the
ronmental samples. Bacterial species found in the
ATU system on Wednesday fan coil air flow were similar to those recovered in
indoor air. In outdoor air, environmental species
such as Aerococcus viridans and T. inchonensis
were mainly encountered.
bacterial contamination inside the building than Considering that to our knowledge, this is
outside. the first time that the BIOLOG system has
been used for airborne bacterial identification,
Microbiological analyses of the humidification this method had a good performance, leading to
water tank the good identification of 94.3% of the analysed
colonies.
Results reported on Table 7 showed that the total
bacterial count in the humidification water tank Fungal identification
was high (range between 1 × 103 CFU per 100 mL
and 1.5 × 105 CFU per 100 mL) with an increasing Figure 2 shows the seasonal distribution of the
trend during warmer seasons (spring and sum- fungal genera in indoor air. According to other
mer). Legionella spp. was never recovered in the studies, Penicillium spp., Aspergillus spp. and
ATU humidification water. Cladosporium cladosporioides and C. sphaeros-
permum were the most widespread fungal gen-
Bacterial identification era identified in indoor air (Pasanen et al. 1992;
Hyvärinen et al. 1993; Nevalainen and Seuri 2005;
The identification of the most widespread bac- Sampò and Luppi Mosca 1988, 1989). Penicillium
teria in indoor air found Micrococcus (32% of was predominant in indoor air during the winter
species) and Staphylococcus (44%) as the most (68%) and spring (60%), while Cladosporium was
frequently occurring genera. These results support the most widespread genus in summer. This trend
the general idea that, in indoor air, the main was similar to that commonly reported in outdoor
represented bacteria are micrococci and staphy- air (Jo and Seo 2005). The fungal genera identi-
lococci (Nevalainen 1989; Maroni et al. 1993). fied (Penicillium, Aspergillus and Cladosporium)
Micrococcus luteus and Staphylococcus haemolyti- in indoor air are recognised as possible causes of
cus were found in all indoor air samples. Species respiratory allergies (Jones 1999), and Penicillium
commonly found on human skin (Micrococcus species can be associated with sick building syn-
lylae, M. luteus and Staphylococcus saprophyti- drome (Schwab and Straus 2004).

Table 7 Microbiological contamination of humidification water tank

Winter Spring Summer Winter2
Bacteria at 37◦ C (CFU/100 mL) 1,900 29,200 43,000 1,800
Bacteria at 22◦ C (CFU/100 mL) 1,100 150,000 136,000 15,600
Legionella spp. (CFU/L) <100a <100a <100a <100a
a<100 CFU/L: detection limit for Legionella spp. analysis
480
Fig. 2 Distribution of
fungal genera in indoor 100%
air during different
seasons
80%
60%
40%
20%
0%
Winter1 Spring
Discussion
This study evaluated the number and composi-
tion of bacteria and fungi in the indoor air of an
Italian office building equipped with an HVAC
system. It was not performed in response to occu-
pant complaints but rather to describe the build-
ing’s air quality over time. As represented by our
data, bacterial and fungal concentrations in all
offices ranged from 50 to 500 CFU/m3 , indicating
a medium–low contamination level (CEC 1993).
The values obtained were in the range reported
in other studies of non-complaint office buildings
around the world (Parat et al. 1997; Law et al.
2001; Sessa et al. 2002; Shelton et al. 2002; Tsai and
Macher 2005). On the other hand, during the first
winter sampling, a high fungal contamination was
recorded (range between 500 and 2,000 CFU/m3 )
in indoor air. This result is probably related to
the flood that occurred during that period, leading
to water stagnation and environmental conditions
(e.g. relative humidity and nutrients) favouring
fungal growth. No significant difference in the
microbial air contamination was found among the
offices, probably because the three offices consid-
ered were located in an open space without any
particular source of microbial contamination. This
homogeneous distribution of bacterial and fungal
counts was also observed in other studies (Parat
et al. 1997).
Considering the bacterial species identified in
indoor air, the predominance of Micrococcus and
Staphylococcus genera could be due to the human
presence. This finding is supported by the indoor/
Environ Monit Assess (2010) 161:473–483
Other
Mycelia sterilia
Cladosporium
Aspergillus
Penicillium
Summer Winter2
outdoor ratio close to 1 for bacteria at 37◦ C
(0.82 ≤ I/O ≤ 1.13). As observed in other studies
(Pastuszka et al. 2000), the high bacterial count
within the building compared to that observed
outdoor could be associated with various internal
sources, including human activities. Otherwise,
the fungal I/O ratio <1 (0.28 ≤ I/O ≤ 0.8) indi-
cated the absence of an indoor air contamination
source. Moreover, as observed by Burge et al.
(2000) and Shelton et al. (2002), the seasonal dis-
tribution of fungal genera in indoor air confirmed
the influence of outdoor air. The data analysis
revealed the absence of any relationship between
bacterial and fungal counts, confirming their ori-
gin from different sources.
Although some bacterial species encountered
in indoor air can be considered to be opportunistic
pathogens (S. haemolyticus, S. epidermidis and
S. hominis), the heterogeneity of the bacterial
species and the low counts recorded do not sug-
gest any particular risk for occupant health. In
contrast, the high fungal contamination and the
presence of potentially allergenic genera (Peni-
cillium) observed in the first winter sampling
(related to the flood) point out a possible health
risk in that situation.
Important information about influence of the
HVAC characteristics (e.g. ATU, fan coil, water)
on IAQ can be obtained from the detailed analysis
of bacterial and fungal counts. In particular, the
significant daily trend of indoor air bacterial (22◦ C
and 37◦ C) and fungal contamination could be as-
sociated with air movement caused by the daily
HVAC system being switched on in the morning
Environ Monit Assess (2010) 161:473–483
and switched off at the end of the working day.
This trend suggests that bacteria and fungi could
proliferate in the ATU filter and vent ducts when
the HVAC system is switched off. Then, when the
HVAC system is switched on, the microorganisms
could spread in the air. The emission of bacteria
and fungi due to incubation during the night-
time HVAC shutdown was also observed in other
studies (Law et al. 2001).
The significant seasonal trend of microbial
counts observed in indoor air, which differed from
outdoor air, was probably due to many factors
such as internal air circulation and the intermit-
tent use of HVAC system, which varied according
to the season. Therefore, as observed by other
authors (Parat et al. 1997; Tsai and Macher 2005),
indoor microbial counts do not seem to be related
to outdoor levels in the presence of low microbial
contamination. The results of the ATU investiga-
tion demonstrated the good efficiency of micro-
bial contamination removal of the unit. Except
for the first winter sampling, in normal climatic
conditions the ATU seems to have removed air
microbial contamination.
A relevant role in air movement might be at-
tributed to building occupants’ personal choice to
switch on or off the fan coil units. The analysis
of the fan coil unit air flow revealed microbial
contamination levels lower than the indoor air.
Moreover, bacterial species found in the fan coil
unit air flow were similar to those recovered in
the indoor air. These results suggest that fan
coil units determined air movement, but in this
case, they were not directly involved in indoor air
contamination.
In regards to the water investigation, high
bacteria in the humidification water tank was
recorded, underlining a microbial proliferation,
especially during warmer seasons.
In conclusion, this study can help better char-
acterise bacterial and fungal populations in the in-
door air of office building, highlighting the role of
the HVAC system on IAQ. The results obtained
suggest that the uninterrupted functioning of the
HVAC system may be recommended, with the
possible application of an energy-saving device.
Moreover, a programmed and periodical clean-
ing operation and maintenance activities of the
HVAC system should be organised as a preven-
481
tive measure. It was also speculated that great care
should be taken during the cleaning operation of
the water tank in the ATU system. Although in
the investigated building there was no particular
risk observed, except for the unusual flood, it is
important to note the usefulness of an IAQ pre-
ventive evaluation to avoid occupant complaints
or health effects. In fact, although the preventive
investigation may be initially more expensive and
time consuming, it may prove to be useful by pre-
venting building environmental problems whose
solution could be very difficult and need greater
investment.
Acknowledgements This study was carried out with fi-
nancial support from the Telecom Italia Laboratories. The
authors wish to thank Prof. Roberto Bono and Dr. Elisa
Gamalero for critical reading of the manuscript.
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